The present invention refers to new strains of the genus Lactobacillus, and an equine feed product comprising one or more of said strains.
Disturbances in the equine intestinal flora can bring about a deteriorated ability to take up the nutriment from the feed, a reduced efficiency, a deteriorated resistance to infections and accompanying infections of different kinds. Other symptoms are a changed consistency of the excrements, which are often malodorous, and an ugly coat of hair. It is well known that disturbances of the intestinal flora above all appear in stress situations, for instance during long transportations or change of stable environment, too tough training and change of feed, but also in treatment with antibiotics. Problems of a disturbed intestinal flora also occur in foals, which during their first living have to obtain an intestinal microflora of their own.
Different disorders which can be associated with a change of the intestinal bacterial flora are inflammations in the intestinal mucosa and gastric ulcer, and bacterial infections giving for instance acute colitis, such as intestinal chlostridiosis, manifesting in diarrhea and toxemia and caused by i.a. Clostridium difficile, and colic, such as Baron-Gruff-disease, which is caused by Clostridium perfringens. 
The presence of coliform bacteria in the excrements has since long been used to judge the state of health in horses as to disturbances of the intestinal flora. A high content of said bacteria has been considered to indicate that the intestines are in good condition, while a low content thereof, as well as a high content of Bacillus and Clostridium spp, respectively, have been associated with intestinal disturbances. The presence of moulds and other fungus also have been considered to have an injurious effect on the intestinal flora. In addition a reduced body temperature, lower than 37xc2x0 C., has been considered to indicate problems in the gastrointestinal tract. The normal temperature in horses is within the range 37.3-37.8xc2x0 C.
Lactobacilli is one of the main groups of microorganisms in the intestinal microflora of healthy horses. Fujisawa et al. discloses in Bifidobacteria Microflora, Vol. 12(2), p. 87-90, 1993, the isolation of 286 strains of lactobacilli from the gastrointestinal tract of healthy horses. The main part of the strains could be identified as belonging to the species L. salivarius, L. acidophilus and L. reuteri and the last two species were found in all parts of the gastrointestinal tract while L. salivarius was not isolated from the stomach. It is also important to remember that the microflora is also influenced by the type of feed that the horse eats.
Probiotics, that is different bacterial and yeast preparations which are orally administrated with the purpose to improve the health in man and animals, have since long been used against intestinal disturbances to improve the intestinal function and the general state of health. As probiotics are commonly used different strains of lactic acid producing bacteria, such as strains of Lactobacillus and Streptococcus. The probiotic hypotesis says that if said bacteria can be added to the intestines in a sufficient amount during conditions of stress or illness, that is when the intestinal flora is disturbed in such a way that pathogenic bacteria are favoured, an establishment of the not wanted bacterial strains in the intestines will be counteracted. To maintain a healthy intestinal flora would then be decisive to prevent those disorders which are associated with a disturbance of the intestinal flora.
In Fox, Veterinary Medicine, August 1988, Probiotics: Intestinal inoculants for production animals, a review is made of different commercial probiotics used in raising different animals, especially chickens, pigs and calves.
It is today common to feed horses having malodorous excrements with sour milk or yoghurt to overcome the underlying disturbances in the stomach and intestines. Equine products on the market are for instance Bakteriebalans (Lantmxc3xa4nnen, Sweden) and Lactosat(copyright) hxc3xa4st (Ewos Sverige AB, Sxc3x6dertxc3xa4lje) which both contain three different strains of lactic acid producing bacteria, that is of Lactobacillus bulgaricus, Streptococcus thermophilus and Streptococcus faecium. Said agents have in common that the lactobacilli which are present in the administrated products either are such which are naturally prevailing in the fermented product or such which have been isolated from man.
Another agent which has been used against equine intestinal disturbances is ColiCure (VETTECH AB, Stockholm), a product consisting of a purified and stabilized strain of Eschericia coli, which was originally isolated from horse, in a concentration of  greater than 109 viable bacteria/ml. The same principle is behind the administration of xe2x80x9cmashxe2x80x9d, that is dispersed excrements from a healthy horse, that is to get a bacterial strain which fits into the intestinal environment and can become established therein. It is, however, controversial to add bacteria of the family Enterobacteriaceae, for instance E. coli, in view of pathogenicity and occurrence of endotoxines.
EP 0 203 586 discloses a composition for treatment of gastrointestinal disorders in animals, especially those being caused by enterotoxigenic strains of Escherichia coli, which is based on the strain Lactobacillus fermentum ATCC 53113, which has been isolated from pig. An advantage of the new composition is said to be that it is not pathogenic and acts by preventing infectious organisms from binding to the intestinal mucosa. An overview is also given of different bacteria which have been used to prevent or to remedy gastro-intestinal disorders, but which have all turned out to be effective only in part.
It has previously been shown that certain strains of Lactobacillus plantarum have the ability for a certain period of time to become established in the intestinal system of man. In EP 92916294.9 certain strains of Lactobacillus, especially L. plantarum 299 DSM 6595 and variants thereof having the ability to become established in the intestines and to remain for at least 11 days after oral administration, are described. Said strains have turned out to have a favourable effect on different disturbances of the intestinal microflora in man.
The normal lactobacilli flora in the intestines of different animal species are different and although the same species of Lactobacillus can be found in two different animal species, the bacteria do not normally belong to the same strain of said species. The strains present differences between themselves which are of importance for their chances of becoming established in the actual environment. In order to be able to treat gastrointestinal disorders in horses, there is therefore a continous need of a safe, non-toxic product comprising one or more bacterial strains having the ability to become established in the equine intestines.
The invention refers to an equine feed product preventing and rectifying microbially caused intestinal disturbances, which contains one or more strains of Lactobacillus which have been isolated from the gastric or intestinal mucosa of horse and which have the ability to colonize in the equine intestines.
Horse refers in this connection to subjects of the genus Equidae, that is in addition to horse also donkey and zebra, having a similar composition of the intestinal flora.
The feed product according to the invention is preferably a feed which has been fermented with a strain of Lactobacillus which has been isolated from the gastric or intestinal mucosa of horse and which has the ability to colonize in the equine intestines.
The feed product according to the invention is preferably based on a cereal based grain or supplementary feed, green forage or hay.
A preferred aspect of the invention is a feed product based on oats which has been fermented with one or more strains of lactobacilli isolated from horse, especially with any of the new strains stated below.
According to another aspect of the invention the feed product is an ensilage based on green forage or hay which has been fermented with one or more strains of lactobacilli isolated from horse.
The invention also refers to the new strain Lactobacillus plantarum JI:1, which has been desposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH on Apr. 21, 1997, under number DSM 11520 and variants thereof; especially for use for prophylaxis or treatment of disturbances in the intestinal microflora.
The invention also refers to another new strain Lactobacillus species AC:3 and variants thereof, which has been deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH on Oct. 2, 1998 under number DSM 12429.
The above strains have turned out to grow well on oats and give a tasty feed product, survive the passage through the gastrointestinal tract of the horse and have the ability to become established in the intestines of horse. Tests have shown that the new strain Lactobacillus plantarum JI:1 reduces the content of undesired bacteria in the equine intestines, for instance the total count of Bacillus and the total count of Clostridium and seems to improve the immune response of the horse.
The feed product according to the invention can be a supplementary feed or a feed, in solid or liquid form. The feed product can be a grain or supplementary feed based on any type of feed which can be fermented by the selected strains of lactobacilli, for instance cereals, such as oats, i.a. black oats, barley, wheat, rye, sorghum or corn, or vegetables, such as leguminous plants, especially soybeans, root vegetables and cabbage, or green forage, such as grass or hay.
Oats is a preferred feed owing to the high content of vitamins and minerals, the well-balanced amino acid composition, the contents of polar lipids and fibres. A variety of oats having a higher fat content has recently turned out to be an appropriate feed for high capacity horses.
A preferred aspect of a feed product according to the invention can be prepared from oats in accordance with the international patent application WO89/08405 referring to the preparation of a nutrient composition. Other examples of preparation of preferred feed products are given below.
The invention also refers to the use of one or more strains of Lactobacillus which have been isolated from the gastric or intestinal mucosa of horse having the ability to colonize in the equine intestines.
The invention also especially refers to the use of the new strain Lactobacillus plantarum JI:1 or a variant thereof for the prophylaxis or treatment of disturbances in the equine intestinal flora. Also inflammatory conditions in the gastric or intestinal mucosa can be alleviated. The invention especially refers to the use of said strain for the preparation of a product for prophylaxis or treatment of disturbances in the equine intestinal flora.
A feed product according to the invention can be administered before or in connection with stress situations, for instance during the warming up before a race, transportation etc. to horses which on occasions like those are stroken by diarrhea; to improve the common condition of horses which are thin, have a poor appetite or a bad coat; to make horses less sensitive to infections, that is to strengthen the immune response; during treatment with anti-biotics to prevent the disturbances of the intestinal flora which normally appear, and thus the diarrhea which is often associated with treatment with antibiotics; to horses having a gastric ulcer; to horses having a diarrhea or malodorous excrements which might be infected with Clostridium difficile or any other pathogenic microorganism.
The content of lactobacilli in the feed should not be below 5xc2x7108 CFU/g or ml and an adequate dose to grown up horses is 2.1012 CFU per day, which corresponds to about 2-4 litres of the oatmeal gruel exemplified below or about 100-200 ml of a freeze culture as below. Higher contents can without any risk be used on special occations. When administrated to fouls a smaller amount should be given.